The QIAscout device together with its core technology, the QIAscout 12,000-Microraft Array, simplifies single-cell isolation while providing cells with a suitable environment similar to a standard cell culture dish. Whether you are performing single-cell sequencing, whole genome and transcriptome amplification or single-cell miRNA analysis; all you need to get started is your cell sample, the QIAscout device and an inverted microscope. The single cells isolated with the QIAscout can be combined with QIAGEN’s single-cell kits for your direct analysis or used for clonal expansion.

Explore the virtual demo to learn more about the QIAscout, see how easy it is to setup and start working, and find a guide to compatible microscopes and objectives.

The QIAscout instrument platform comprises a controller, power supply for controller, a magnetic collection plate, a motorized release device, 4 inserts for release device, protective lid for release device and a magnetic wand. The instrument can only work in conjunction with the QIAscout arrays.

The QIAscout consists of the QIAscout instrument platform comprising a controller, power supply for controller, a magnetic collection plate, a motorized release device, 4 inserts for release device, protective lid for release device and a magnetic wand as well as a starter pack of 5 QIAscout 12,000-Microraft Arrays

No cell contamination observed with the magnetic wand in Pyrosequencing analysis.

A Representative pyrogram showing the G>A point mutation in the human EGFR gene in a sample with single SW48 cells (positive control). B Representative pyrogram showing absence of the A allele in single HT-29 cells, transferred with the same unwashed magnetic wand as the SW48 cells. C Frequency of the A allele in two pierced HT-29 cells (negative control), two pierced SW48 cells (positive control) and 24 pierced HT-29 cells. Single SW48 cells transferred using a clean magnetic wand (positive control) show a mean frequency of A allele (=point mutation) of 50.7%. Pierced single HT-29 cells transferred using an unwashed magnetic wand and analyzed for the presence of SW48 cell contamination showed a similar frequency of A allele when compared to the pierced single HT-29 cells transferred using the clean magnetic wand (negative control), indicating no contamination between the two cell lines, the arrays and the microrafts.

No DNA contamination observed with the release needle in qPCR analysis.

Cells remain viable for several days and form subpopulations, irrespective of whether the pierced microrafts containing single cells are transferred to a cell culture dish or single cells present on microrafts are left intact and allowed to grow on the QIAscout array itself (see corresponding Application Note titled “Rapid and Efficient Method of Isolation and Recovery of Highly Viable Single Cells Free of Contamination” under the “Product Resources” tab).

The QIAscout method allows efficient and reliable isolation of single targeted cells. Selection of cells can be based on different cellular parameters like cell morphology or fluorescence. Conventional manual methods like pipetting and serial dilution are less successful in selective isolation of single cells when compared to the novel QIAscout technology (data not shown).

The QIAscout system is an effective and fast method to isolate viable and pure single cells, ensuring minimal manipulation of the cellular status. The core technology of the QIAscout is the QIAscout array, which is composed of 12,000 individual microrafts. Each microraft is 200 µm in length and 200 µm in width and is securely held within a micron-scale well of the QIAscout array. Within the QIAscout array, the microrafts serve as releasable culture sites for individual cells or colonies.

The QIAscout array is ideal for various cell types including adherent cells, suspension cells, cell lines, primary cells, and fluorescent cells providing an environment suitable for their growth and viability similar to any standard cell culture dish. QIAscout arrays are mounted on inverted microscopes and can be imaged by brightfield, fluorescence and confocal microscopy.

A motorized release device is mounted onto a 4x, 5x or 10x objective of common inverted lab microscopes to allow for selection and picking of individual microrafts. To isolate target cells, simply press the button on the controller which makes the release needle pierce through the lower surface of the QIAscout array and dislodge an individual microraft with your cell of interest attached. The released microraft is then recovered using a magnetic wand and transported to a secondary vessel (e.g., reaction tube of downstream application).

Processing the collected cell for downstream application or further cultivation

The cell distribution on the QIAscout array follows a Poisson distribution (see Table “Cell distribution on the array follows a Poisson distribution”). To achieve the maximum number of microrafts on the QIAscout array containing a single cell and to minimize the number of microrafts with no cell or more than one cell, it is suggested to plate cells at a cell : microraft ratio of 1:2 or 1:3. A QIAscout array contains 12,000 individual microrafts, and therefore plating 4,000–6,000 cells is generally recommended. Adjust the cell density of your suspension and add the desired number of cells to the QIAscout array in 2 ml of medium.

Cell distribution on the array follows a Poisson distribution

Cell : Well ratio

% Empty wells

% Wells capturing a single cell

% Wells capturing 2 or more cells

1:1

37

37

26

1:2

61

30

9

1:3

72

24

4

1:5

82

16

2

1:10

90

9

1

After adding cells to the QIAscout array, allow the cells to settle and wait until the cells have attached firmly enough to the microraft surface to allow for release and collection. A waiting time of 6–10 hours in culture is generally adequate, but the length of this period will be cell line-dependent. Once the cells are adhered to the microrafts, the QIAscout array can be screened under an inverted microscope to identify target cells and colonies and proceed with isolation and recovery.

Applications

The QIAscout is highly suitable for laboratories performing single-cell research. Single-cell isolation with QIAscout is compatible with multiple downstream applications such as:

Whole Genome Amplification

Whole Transcriptome Amplification

Single-cell Sequencing

Single-cell miRNA Analysis

Clonal Expansion

Features

Specifications

Altitude

Up to 2000 m (6500 ft.)

Applications

Single-cell isolation

Connections

Connect motor cable connector of the release device onto the controller until a snap-click sound is heard. To disconnect the motor cable, hold on to the connector and pull out the cable.